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We narrowed to 994 results for: Des

Showing: 881 - 900 of 994 results
  1. Gibson Assembly Protocol

    Type
    Protocol
    ... Procedure Design your plasmid and order primers (see figure to the right). When designing your plasmid...and enrich for correctly assembled plasmids by designing primers to split an antibiotic resistance gene...and yield. If there are significant amounts of undesired product, gel-purify DNA segments. Otherwise, PCR... High-Fidelity DNA Polymerase - incorporates nucleotides to “fill in” the gaps in the annealed DNA fragments... (<150 bp). Pro-Tip Please note that the way to design the “stitching” primers and the amounts of primers...window) . NEB has other resources, such as a primer design tool (Link opens in a new window) . Gibson Assembly...
  2. Lab Safety for Biosafety Levels One and Two

    Type
    Protocol
    ...There are four biosafety levels. This protocol provides information for both biosafety level 1 (BSL-1)...level has different safety requirements. BSL-1 is designated for those working with microbes that don’t cause...Staphylococcus aureus or Vibrio cholerae . BSL-2 includes all of the precautions needed in BSL-1, along ...before and after working in the lab. Ensure that a designated chemical waste accumulation site is present, ...(MSDS). While You Work Dispose of sharps in a designated sharps container. Sharps include anything that... drain. Solid BSL-2 waste can be collected in designated biohazardous waste containers that can be autoclaved...important! Disclaimer: The SOP presented here has been designed by the Addgene nonprofit plasmid repository and...
  3. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...created. Additional design rules can be found at the Broad Institute's TRC shRNA Design Process . To minimize...Consortium A.2 Map of pLKO.1 A.3 Related plasmids B. Designing shRNA Oligos for pLKO.1 B.1 Determine the optimal...sites in place of the stuffer. The AgeI site is destroyed in most cases (depending on the target sequence....9kb stuffer, visit www.addgene.org/10878/ . Description Vector Element U6 Human U6 promoter drives RNA...the pLKO.1 TRC-cloning vector. Addgene Plasmid Description pLKO.1 – TRC control Negative control vector ...website and “search for “pLKO”“. Back to Top B. Designing shRNA Oligos for pLKO.1 B.1 Determining the Optimal...accessed after a free registration. Guidelines for designing siRNAs with effective gene silencing: Starting...
  4. Antibodies 101: Fab Fragments

    Type
    Blog Post
    ...talking about here? Many of the Fab fragments described above can be used for similar applications and...
  5. RNA Extraction Without A Kit

    Type
    Blog Post
    ...Allewell and Sarma, 1974). Additionally, ThermoFisher provides a protocol on how to integrate RNAlater® with ...
  6. Working with Nuclear Receptors

    Type
    Blog Post
    ...Activation of membrane receptors and signaling cascades conceivably allows the genome to sense the impact...
  7. Plasmid Cloning by Restriction Enzyme Digest (with Protocols)

    Type
    Protocol
    ...restriction enzyme digest (subcloning), including design and experimental procedures....easily move YGOI into a mammalian expression vector. Design (Choosing enzymes) Many DNA analysis tools, including..., but do not cut within your insert Are in the desired location in your recipient plasmid (usually in ...your recipient plasmid as well as a specifically designed test digest later to verify that the insert was... fear. You have other options, such as: Adding desired restriction sites to flank your insert : You can...restriction enzymes that cut within your insert. Adding desired restriction sites to your recipient plasmid : You... orientation of your insert, so you may want to design a diagnostic digest for this purpose. Congratulations...
  8. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...Vector Guides Virus Blog Posts Addgene Protocols Viral Service Introduction This protocol describes how ...This protocol describes how to generate a monoclonal cell line from a polyclonal pool of stable cells...type will be able to grow under the conditions described in this protocol. Conditioned medium (details ... specific conditions used. Each 96-well plate described in this protocol could, in theory, give rise to...protocol was performed at Addgene on the A549 cells described in our sample data , each 96-well plate gave rise...from Feng Zhang (Addgene plasmid #52962 ) and is described in Improved vectors and genome-wide libraries ...
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